Posttranslational Regulation of Ty1 Retrotransposition by Mitogen-Activated Protein Kinase Fus3

ABSTRACT

Ty1 retrotransposons in Saccharomyces cerevisiae are maintained in a state of transpositional dormancy. We isolated a mutation, rtt100-1, that increases the transposition of genomic Ty1 elements 18- to 56-fold but has little effect on the transposition of related Ty2 elements. rtt100-1 was shown to be a null allele of the FUS3 gene, which encodes a haploid-specific mitogen-activated protein kinase. In fus3mutants, the levels of Ty1 RNA, protein synthesis, and proteolytic processing were not altered relative to those in FUS3strains but steady-state levels of TyA, integrase, and reverse transcriptase proteins and Ty1 cDNA were all increased. These findings suggest that Fus3 suppresses Ty1 transposition by destabilizing viruslike particle-associated proteins. The Fus3 kinase is activated through the mating-pheromone response pathway by phosphorylation at basal levels in naive cells and at enhanced levels in pheromone-treated cells. We demonstrate that suppression of Ty1 transposition in naive cells requires basal levels of Fus3 activation. Substitution of conserved amino acids required for activation of Fus3 derepressed Ty1 transposition. Moreover, epistasis analyses revealed that components of the pheromone response pathway that act upstream of Fus3, including Ste4, Ste5, Ste7, and Ste11, are required for the posttranslational suppression of Ty1 transposition by Fus3. The regulation of Ty1 transposition by Fus3 provides a haploid-specific mechanism through which environmental signals can modulate the levels of retrotransposition.

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Posttranslational Regulation of Ty1 Retrotransposition by Mitogen-Activated Protein Kinase Fus3

ACKNOWLEDGMENTS

We are grateful to B. Faiola and W.-Y. Hu for their assistance at an early stage in this project, to H. Madhani for helpful suggestions and for sharing results prior to publication, to M. Bryk and R. Zitomer for helpful comments on the manuscript, and to the Molecular Genetics Core Facility for oligomer synthesis.

This work was funded by National Institutes of Health grant GM52072 to M.J.C. The research of D.J.G. was sponsored by the National Cancer Institute, DHHS, under contract with ABL.